In biomedical imaging, all optical techniques face a fundamental trade-off between spatial resolution and tissue penetration. Therefore, obtaining an organelle-level resolution image of a whole organ has remained a challenging and yet appealing scientific pursuit. Over the past decade, optical microscopy assisted by mechanical sectioning or chemical clearing of tissue has been demonstrated as a powerful technique to overcome this dilemma, one of particular use in imaging the neural network. However, this type of techniques needs lengthy special preparation of the tissue specimen, which hinders broad application in life sciences. Here, we propose a new label-free three-dimensional imaging technique, named microtomy-assisted photoacoustic microscopy (mPAM), for potentially imaging all biomolecules with 100% endogenous natural staining in whole organs with high fidelity. We demonstrate the first label-free mPAM, using UV light for label-free histology-like imaging, in whole organs (e.g., mouse brains), most of them formalin-fixed and paraffin- or agarose-embedded for minimal morphological deformation. Furthermore, mPAM with dual wavelength illuminations is also employed to image a mouse brain slice, demonstrating the potential for imaging of multiple biomolecules without staining. With visible light illumination, mPAM also shows its deep tissue imaging capability, which enables less slicing and hence reduces sectioning artifacts. mPAM could potentially provide a new insight for understanding complex biological organs.
Terence T. W. Wong, Ruiying Zhang, Hsun-Chia Hsu, Konstantin I. Maslov, Junhui Shi, Ruimin Chen, K. Kirk Shung, Qifa Zhou, and Lihong V. Wang, "Whole-organ atlas imaged by label-free high-resolution photoacoustic microscopy assisted by a microtome," Proc. SPIE 10494, Photons Plus Ultrasound: Imaging and Sensing 2018, 104942N (Presented at SPIE BiOS: January 31, 2018; Published: 19 February 2018); https://doi.org/10.1117/12.2291056.
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